当归红芪超滤物对过氧化氢致内皮细胞凋亡中热休克蛋白70及内皮型一氧化氮合酶表达的影响

摘要

Objective To investigate the effects of ultra-filtration extract from the mixture of Angelicae Sinensis Radix and Hedysarum Polybotrys (UFE-AH) on the expressions of HSP70 and eNOS in H2O2-induced endothelial cell apoptosis. Methods H2O2 induced ECV-304 cell apoptosis to prepare models. The experiment was divided into normal control group, model group, simple medicine group, medicine intervention group, and all treatment groups received relevant medicine for intervention. Flow cytometry (FCM) was used to detect apoptosis and concentration of intracellular Ca2+;RT-PCR was used to detect the mRNA expression of HSP70 and eNOS;Western blot was used to detect the expression of HSP70 protein;Nitrale reduetase and spectrophotometric method were employed to detect the content of NO. Results Compared with normal control group, cell apoptosis rate, concentration of intracellular Ca2+, and expression of HSP70 increased significantly in model group (P<0.05);gene expression of eNOS mRNA and content of NO decreased (P<0.05). Compared with model group, cell apoptosis rate and concentration of intracellular Ca2+dropped in medicine intervention group (P<0.05);expressions of HSP70, eNOS mRNA and content of NO increased (P<0.05). Conclusion UFE-AH can confront H2O2-induced cell apoptosis H2O2 of ECV-304 human umbilical vein endothelial by increasing the expressions of HSP70 and eNOS and content of NO, and reducing the intracellular calcium overload.%目的：探讨当归红芪超滤物对过氧化氢(H2O2)致内皮细胞凋亡中热休克蛋白70(HSP70)及内皮型一氧化氮合酶(eNOS)表达的影响。方法 H2O2诱导 ECV-304人脐静脉内皮细胞凋亡制备模型,实验分为正常对照组、模型组、单纯药物组、药物干预组,并给予相应药物干预,流式细胞仪检测细胞凋亡及细胞内 Ca2+浓度,RT-PCR法检测HSP70、eNOS的基因表达,蛋白印迹法检测HSP70的蛋白表达,硝酸还原酶法及分光光度法检测一氧化氮(NO)的含量。结果与正常对照组比较,模型组细胞凋亡率明显增加、细胞内Ca2+浓度明显上升(P＜0.05),细胞HSP70基因和蛋白表达上调、eNOS基因表达下调及NO含量降低(P＜0.05)；与模型组比较,药物干预组细胞凋亡率、细胞内Ca2+浓度明显降低(P＜0.05),细胞HSP70基因和蛋白表达上调、eNOS基因表达上调及NO含量升高(P＜0.05)。结论当归红芪超滤物通过上调HSP70、eNOS的表达及NO含量,降低细胞内钙超载,发挥抗H2O2致ECV-304人脐静脉内皮细胞凋亡的作用。